Mixtures and compositions comprising Paenibacillus strains or metabolites thereof and other biopesticides

Abstract

The present invention relates to novel mixtures comprising, as active components, at least one isolated bacterial strain, which is a member of the genus Paenibacillus, or a cell-free extract thereof or at least one metabolite thereof, and at least one other biopesticide. The present invention also relates to compositions comprising at least one of such bacterial strains, whole culture broth or a cell-free extract or a fraction thereof or at least one metabolite thereof, and at least one other biopesticide. The present invention also relates to a method of controlling or suppressing plant pathogens or preventing plant pathogen infections by applying such composition. The present invention also relates to mixtures of fusaricidins which are pesticidal metabolites produced by the abovementioned strains, and other biopesticides.

The invention claimed is: 
     
       1. A mixture, comprising as active components
 1) at least one  Paenibacillus  strain or a culture medium thereof, a cell-free extract thereof, or at least one metabolite thereof; 
 wherein the at least one  Paenibacillus  strain is selected from the group consisting of: 
 a)  Paenibacillus  strain Lu16774 deposited with DSMZ under Accession No. DSM 26969; 
 b)  Paenibacillus  strain Lu17007 deposited with DSMZ under Accession No. DSM 26970; 
 c)  Paenibacillus  strain Lu17015 deposited with DSMZ under Accession No. DSM 26971; and 
 d) a  Paenibacillus  strain which comprises a DNA sequence exhibiting
 d1) at least 99.6% nucleotide sequence identity to the DNA sequences SEQ ID NO:4 or SEQ ID NO:9; or 
 d2) at least 99.8% nucleotide sequence identity to the DNA sequence SEQ ID NO:14; or 
 d3) at least 99.9% nucleotide sequence identity to the DNA sequences SEQ ID NO:5 or SEQ ID NO:10; or 
 d4) at least 99.2% nucleotide sequence identity to the DNA sequence SEQ ID NO:15; or 
 d5) at least 99.2% nucleotide sequence identity to the DNA sequences SEQ ID NO:6 or SEQ ID NO:11; or 
 d6) at least 99.8% nucleotide sequence identity to the DNA sequence SEQ ID NO:16; or 
 d7) at least 99.8% nucleotide sequence identity to the DNA sequences SEQ ID NO:7 or SEQ ID NO:12; or 
 d8) at least 99.3% nucleotide sequence identity to the DNA sequence SEQ ID NO:17; or 
 d9) 100.0% nucleotide sequence identity to the DNA sequences SEQ ID NO:8 or SEQ ID NO:13; or 
 d10) at least 99.9% nucleotide sequence identity to the DNA sequence SEQ ID NO:18; 
 
 and 
 2) at least one biopesticide II selected from the groups L1) to L5):
 L1) at least one microbial pesticide with fungicidal, bactericidal, viricidal and/or plant defense activator activity selected from:  Ampelomyces quisqualis, Aspergillus flavus, Aureobasidium pullulans, Bacillus altitudinis, B. amyloliquefaciens, B. megaterium, B. mojavensis, B. mycoides, B. pumilus, B. simplex, B. solisalsi, B. subtilis, B. subtilis  var.  amyloliquefaciens, Candida oleophila, C. saitoana, Clavibacter michiganensis  (bacteriophages),  Coniothyrium minitans, Cryphonectria parasitica, Cryptococcus albidus, Dilophosphora alopecuri, Fusarium oxysporum, Clonostachys rosea  f.  catenulate  (also named  Gliocladium catenulatum ),  Gliocladium roseum, Lysobacter antibioticus, L. enzymogenes, Metschnikowia fructicola, Microdochium dimerum, Microsphaeropsis ochracea, Muscodor albus, Paenibacillus alvei, Paenibacillus polymyxa, Pantoea vagans, Penicillium bilaiae, Phlebiopsis gigantea, Pseudomonas  sp.,  Pseudomonas chloraphis, Pseudozyma flocculosa, Pichia anomala, Pythium oligandrum, Sphaerodes mycoparasitica, Streptomyces griseoviridis, S. lydicus, S. violaceusniger, Talaromyces flavus, Trichoderma asperelloides, T. asperellum, T. atroviride, T. fertile, T. gamsii, T. harmatum, T. harzianum, T. polysporum, T. stromaticum, T. virens, T. viride, Typhula phacorrhiza, Ulocladium oudemansii, Verticillium  dahlia, and/or zucchini yellow mosaic virus (avirulent strain); 
 L2) at least one biochemical pesticide with fungicidal, bactericidal, viricidal and/or plant defense activator activity selected from: harpin protein and/or  Reynoutria sachalinensis  extract; 
 L3) at least one microbial pesticide with insecticidal, acaricidal, molluscidal and/or nematicidal activity selected from:  Agrobacterium radiobacter, Bacillus cereus, B. firmus, B. thuringiensis, B. thuringiensis  ssp.  aizawai, B. t . ssp.  israelensis, B. t . ssp.  galleriae, B. t . ssp.  kurstaki, B. t . ssp.  tenebrionis, Beauveria bassiana, B. brongniartii, Burkholderia  spp.,  Chromobacterium subtsugae, Cydia pomonella  granulovirus (CpGV),  Cryptophlebia leucotreta  granulovirus (CrleGV),  Flavobacterium  spp.,  Helicoverpa armigera  nucleopolyhedrovirus (HearNPV),  Helicoverpa zea  nucleopolyhedrovirus (HzNPV),  Helicoverpa zea  single capsid nucleopolyhedrovirus (HzSNPV),  Heterorhabditis bacteriophora, Isaria fumosorosea, Lecanicillium longisporum, L. muscarium, Metarhizium anisopliae, Metarhizium anisopliae  var.  anisopliae, M. anisopliae  var.  acridum, Nomuraea rileyi, Paecilomyces fumosoroseus, P. lilacinus, Paenibacillus popilliae, Pasteuria  spp.,  P. nishizawae, P. penetrans, P. ramosa, P. thornea, P. usgae, Pseudomonas fluorescens, Spodoptera littoralis  nucleopolyhedrovirus (SpliNPV),  Steinernema carpocapsae, S. feltiae, S. kraussei, Streptomyces galbus , and/or  S. microflavus;    
 L4) at least one biochemical pesticide with insecticidal, acaricidal, molluscidal, pheromone and/or nematicidal activity selected from: L-carvone, citral, (E,Z)-7,9-dodecadien-1-yl acetate, ethyl formate, (E,Z)-2,4-ethyl decadienoate (pear ester), (Z,Z,E)-7,11,13-hexadecatrienal, heptyl butyrate, isopropyl myristate, lavanulyl senecioate, cis-jasmone, 2-methyl 1-butanol, methyl eugenol, cis-jasmone, methyl jasmonate, (E,Z)-2,13-octadecadien-1-ol, (E,Z)-2,13-octadecadien-1-ol acetate, (E,Z)-3,13-octadecadien-1-ol, R-1-octen-3-ol, pentatermanone, (E,Z,Z)-3,8,11-tetradecatrienyl acetate, (Z,E)-9,12-tetradecadien-1-yl acetate, Z-7-tetradecen-2-one, Z-9-tetradecen-1-yl acetate, Z-11-tetradecenal, Z-11-tetradecen-1-01, extract of  Chenopodium  ambrosiodes, Neem oil, and/or Quillay extract; 
 L5) at least one microbial pesticide with plant stress reducing, plant growth regulator, plant growth promoting and/or yield enhancing activity selected from:  Azospirillum amazonense, A. brasilense, A. lipoferum, A. irakense, A. halopraeferens, Bradyrhizobium  spp.,  B. elkanii, B. japonicum, B. liaoningense, B. lupini, Delftia acidovorans, Glomus intraradices, Mesorhizobium  spp.,  Rhizobium leguminosarum  by.  phaseoli, R. I . bv.  trifolii, R. I . bv.  viciae, R. tropici , and/or  Sinorhizobium meliloti,    
 
 wherein the mixture comprises component 1) and component 2) in a synergistically effective amount. 
 
     
     
       2. The mixture of  claim 1 , wherein said at least one  Paenibacillus  strain has antifungal activity against at least two of the plant pathogens selected from the group consisting of  Alternaria  spp.,  Botrytis cinerea, Phytophthora infestans  and  Sclerotinia sclerotiorum.    
     
     
       3. The mixture of  claim 1 , wherein said at least one  Paenibacillus  strain is capable of producing at least one of the following compounds: 
       
         
           
           
               
               
           
         
         in a growth medium comprising at least one source of carbon and one source of nitrogen. 
       
     
     
       4. The mixture of  claim 1 , wherein the component 1) comprises a substantially purified culture of said at leastone  Paenibacillus  strain as defined in  claim 1 . 
     
     
       5. The mixture of  claim 1 , wherein the component 1) comprises a whole culture broth, a culture medium or a cell-free extract of said at least comprises a whole culture broth, a culture medium or a cell-free extract of at least one  Paenibacillus  strain as defined in  claim 1 . 
     
     
       6. The mixture of  claim 1 , wherein the component 1) comprises said at least one metabolite of the at least one  Paenibacillus  strain selected from the group consisting of polymyxins, octapeptins, polypeptins, pelgipeptins and fusaricidins. 
     
     
       7. The mixture of  claim 6 , wherein the at least one metabolite is selected from fusaricidins of formula I 
       
         
           
           
               
               
           
         
         wherein 
         R is selected from the group consisting of 15-guanidino-3-hydroxypentadecanoic acid (GHPD) and 12-guanidinododecanoic acid (12-GDA); 
         X 1  is threonine; 
         X 2  is selected from the group consisting of isoleucine and valine; 
         X 3  is selected from the group consisting of tyrosine, valine, isoleucine and phenylalanine; 
         X 4  is threonine; 
         X 5  is selected from the group consisting of glutamine and asparagine; 
         X 6  is alanine; and 
         wherein an arrow defines a single (amide) bond either between the carbonyl moiety of R and the amino group of the amino acid X 1  or between the carbonyl group of one amino acid and the amino group of a neighboring amino acid, wherein the tip of the arrow indicates the attachment to the amino group of said amino acid X 1  or of said neighboring amino acid; and 
         wherein the single line without an arrow head defines a single (ester) bond between the carbonyl group of X 6  and the hydroxyl group of X 1 . 
       
     
     
       8. The mixture of  claim 7 , wherein the at least one metabolite is selected from fusaricidins of formula I 
       
         
           
           
               
               
           
         
         wherein 
         R is selected from the group consisting of 15-guanidino-3-hydroxypentadecanoic acid (GHPD) and 12-guanidinododecanoic acid (12-GDA); 
         X 1  is threonine; 
         X 2  is isoleucine; 
         X 3  is tyrosine; 
         X 4  is threonine; 
         X 5  is selected from the group consisting of glutamine and asparagine; 
         X 6  is alanine; and 
         wherein an arrow defines a single (amide) bond either between the carbonyl moiety of R and the amino group of the amino acid X 1  or between the carbonyl group of one amino acid and the amino group of a neighboring amino acid, wherein the tip of the arrow indicates the attachment to the amino group of said amino acid X 1  or of said neighboring amino acid; and 
         wherein the single line without an arrow head defines a single (ester) bond between the carbonyl group of X 6  and the hydroxyl group of X 1 . 
       
     
     
       9. The mixture of  claim 8 , wherein the at least one metabolite is selected from fusaricidins 1A and 1 B: 
       
         
           
           
               
               
           
         
       
     
     
       10. A composition comprising the mixture of  claim 1  and an auxiliary. 
     
     
       11. The composition of  claim 10 , further comprising a pesticide III selected from the groups SF) and SI):
 SF) Fungicides
 inhibitors of complex III at Q o  site selected from the group consisting of: pyraclostrobin, azoxystrobin, picoxystrobin, trifloxystrobin, dimoxystrobin, enestroburin, fenaminstrobin, fluoxastrobin, kresoxim-methyl, mandestrobine, metominostrobin, orysastrobin, pyrametostrobin, and pyraoxystrobin; 
 broad-spectrum pyridine and pyrazole inhibitors of complex II selected from the group consisting of: fluxapyroxad, boscalid, benzovindiflupyr, penflufen, penthiopyrad, sedaxane, fluopyram, bixafen, and isopyrazam; 
 Basidiomycetes-specific inhibitors of complex II selected from the group consisting of: carboxin, benodanil, fenfuram, flutolanil, furametpyr, mepronil, oxycarboxin, and thifluzamide; 
 
 ATP production inhibitor silthiofam;
 fungicidal azole compounds selected from the group consisting of: ipconazole, difenoconazole, prothioconazole, prochloraz, triticonazole, flutriafol, cyproconazole, diniconazole, diniconazole-M, fluquinconazole, flusilazole, hexaconazole, imazalil, imibenconazole, metconazole, myclobutanil, simeconazole, tebuconazole, triadimenol, uniconazole, thiabendazole; 
 Oomycetes fungicides selected from: oxathiapiprolin, valifenalate, metalaxyl, metalaxyl-M, ethaboxam, dimethomorph, zoxamide, flumorph, mandipropamid, pyrimorph, benthiavalicarb, and iprovalicarb; 
 MAP/histidine kinase inhibitor: fludioxonil; 
 benzimidazole compounds selected from the group consisting of: 
 
 thiophanate-methyl, and carbendazim;
 dithiocarbamate compounds selected from the group consisting of: thiram, and ziram; 
 
 SI) Insecticides
 GABA antagonist compounds selected from the group consisting of: fipronil, ethiprole, vaniliprole, pyrafluprole, pyriprole, and 5-amino-1-(2,6-dichloro-4-methyl-phenyl)-4-sulfinamoyl-1H-pyrazole-3-carbothioic acid amide; 
 lepidopteran-specific ryanodine receptor inhibitors selected from the group consisting of: chlorantraniliprole and flubendiamide; 
 cross-spectrum ryanodine receptor inhibitor: cyantraniliprole; 
 pyrethroid sodium channel modulators selected from the group consisting of: 
 
 tefluthrin, bifenthrin, cypermethrin, alpha-cypermethrin, cyfluthrin, beta-cyfluthrin, lambda-cyhalothrin, deltamethrin, esfenvalerate, etofenprox, fenvalerate, flucythrinate, and permethrin;
 systemically-active neonicotinoid compounds: clothianidin, imidacloprid, thiamethoxam, dinotefuran, acetamiprid, flupyradifurone, thiacloprid, triflumezopyrim, nitenpyram; 
 Acetylcholinesterase inhibitors, chloride channel activators and sulfoximines: sulfoxaflor, acephate, chlorpyrifos, thiodicarb, abamectin, spinosad; 
 other insecticide: tioxazafen. 
 
 
     
     
       12. A plant propagation material having a coating comprising the composition of  claim 10 . 
     
     
       13. A method of controlling, suppressing pathogens or preventing pathogen infection, wherein the pathogens, their habitat or the materials or plants to be protected against pathogen attack, or the soil or propagation material are treated with an effective amount of the mixture of  claim 1 . 
     
     
       14. A method of controlling, suppressing pathogens or preventing pathogen infection, wherein the pathogens, their habitat or the materials or plants to be protected against pathogen attack, or the soil or propagation material are treated with an effective amount of the composition of  claim 10 .